GSBS Dissertations and Theses

ORCID ID

0000-0003-2084-5871

Approval Date

5-15-2017

Document Type

Doctoral Dissertation

Academic Program

Interdisciplinary Graduate Program

Department

Microbiology and Physiological Systems

First Thesis Advisor

Allan Jacobson, Ph.D.

Keywords

NMD, Decapping, dcp2, dcp1, xrn1, upf

Abstract

mRNA decay is an important step in gene regulation, environmental responsiveness, and mRNA quality control. One such quality control pathway, Nonsense-mediated mRNA Decay (NMD), targets transcripts whose translation terminates prematurely. However, the scope and the defining features of NMD-targeted transcripts remain elusive. To address these issues, we re-evaluated the genome-wide expression of annotated transcripts in yeast cells harboring deletions of the UPF1, UPF2, or UPF3 genes. The vast majority of NMD-regulated transcripts are normal-looking protein-coding mRNAs. Our bioinformatics analyses reveal that this set of NMD-regulated transcripts generally have lower translational efficiency, lower average codon optimality scores, and higher ratios of out-of-frame translation.

General mRNA decay is predominantly mediated by decapping by the Dcp1-Dcp2 complex and 5' to 3' decay by Xrn1, but the exact mechanism of decapping regulation has remained largely unknown. Several in vitro and in vivo studies have revealed the importance of the C-terminal extension of Dcp2 and the identities of many decapping regulators that interact with the decapping complex. To better understand how decapping regulation is achieved by the C-terminal extension of Dcp2 we generated RNA-Seq libraries from a Dcp2 allele that lacks this portion of Dcp2 along with libraries from strains that contain single deletions of several decapping activators. Our transcriptome-wide results indicate that the C-terminal extension of Dcp2 is crucial for efficient regulation of decapping, and different decapping activators are responsible for targeting different sets of mRNAs. Considering the limited pool of Dcp1-Dcp2 in the cell decapping activators might be in competition for decapping complex binding.

Collectively, our results yield valuable insights into the mechanism of substrate selection for mRNA quality control and decay in yeast.

DOI

10.13028/M2H38V

Rights and Permissions

Licensed under a Creative Commons license

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This work is licensed under a Creative Commons Attribution 4.0 License.

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